[English] 日本語
Yorodumi- EMDB-26461: cryo-EM structure of the ADP state wild type myosin-15-F-actin co... -
+Open data
-Basic information
Entry | Database: EMDB / ID: EMD-26461 | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Title | cryo-EM structure of the ADP state wild type myosin-15-F-actin complex (symmetry expansion) | ||||||||||||
Map data | |||||||||||||
Sample |
| ||||||||||||
Keywords | myosin motor proteins / actin cytoskeleton / stereocilia / deafness / MOTOR PROTEIN | ||||||||||||
Biological species | Gallus gallus (chicken) / Mus musculus (house mouse) | ||||||||||||
Method | single particle reconstruction / cryo EM / Resolution: 3.9 Å | ||||||||||||
Authors | Gong R / Reynolds MJ / Alushin GM | ||||||||||||
Funding support | United States, 3 items
| ||||||||||||
Citation | Journal: Sci Adv / Year: 2022 Title: Structural basis for tunable control of actin dynamics by myosin-15 in mechanosensory stereocilia. Authors: Rui Gong / Fangfang Jiang / Zane G Moreland / Matthew J Reynolds / Santiago Espinosa de Los Reyes / Pinar Gurel / Arik Shams / James B Heidings / Michael R Bowl / Jonathan E Bird / Gregory M Alushin / Abstract: The motor protein myosin-15 is necessary for the development and maintenance of mechanosensory stereocilia, and mutations in myosin-15 cause hereditary deafness. In addition to transporting actin ...The motor protein myosin-15 is necessary for the development and maintenance of mechanosensory stereocilia, and mutations in myosin-15 cause hereditary deafness. In addition to transporting actin regulatory machinery to stereocilia tips, myosin-15 directly nucleates actin filament ("F-actin") assembly, which is disrupted by a progressive hearing loss mutation (p.D1647G, ""). Here, we present cryo-electron microscopy structures of myosin-15 bound to F-actin, providing a framework for interpreting the impacts of deafness mutations on motor activity and actin nucleation. Rigor myosin-15 evokes conformational changes in F-actin yet maintains flexibility in actin's D-loop, which mediates inter-subunit contacts, while the mutant locks the D-loop in a single conformation. Adenosine diphosphate-bound myosin-15 also locks the D-loop, which correspondingly blunts actin-polymerization stimulation. We propose myosin-15 enhances polymerization by bridging actin protomers, regulating nucleation efficiency by modulating actin's structural plasticity in a myosin nucleotide state-dependent manner. This tunable regulation of actin polymerization could be harnessed to precisely control stereocilium height. | ||||||||||||
History |
|
-Structure visualization
Supplemental images |
---|
-Downloads & links
-EMDB archive
Map data | emd_26461.map.gz | 282.9 MB | EMDB map data format | |
---|---|---|---|---|
Header (meta data) | emd-26461-v30.xml emd-26461.xml | 17.3 KB 17.3 KB | Display Display | EMDB header |
FSC (resolution estimation) | emd_26461_fsc.xml | 18.1 KB | Display | FSC data file |
Images | emd_26461.png | 47.2 KB | ||
Masks | emd_26461_msk_1.map | 512 MB | Mask map | |
Filedesc metadata | emd-26461.cif.gz | 4.9 KB | ||
Others | emd_26461_half_map_1.map.gz emd_26461_half_map_2.map.gz | 410.3 MB 410.2 MB | ||
Archive directory | http://ftp.pdbj.org/pub/emdb/structures/EMD-26461 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-26461 | HTTPS FTP |
-Validation report
Summary document | emd_26461_validation.pdf.gz | 891.1 KB | Display | EMDB validaton report |
---|---|---|---|---|
Full document | emd_26461_full_validation.pdf.gz | 890.7 KB | Display | |
Data in XML | emd_26461_validation.xml.gz | 26.1 KB | Display | |
Data in CIF | emd_26461_validation.cif.gz | 34.5 KB | Display | |
Arichive directory | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-26461 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-26461 | HTTPS FTP |
-Related structure data
-Links
EMDB pages | EMDB (EBI/PDBe) / EMDataResource |
---|---|
Related items in Molecule of the Month |
-Map
File | Download / File: emd_26461.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Voxel size | X=Y=Z: 1.03 Å | ||||||||||||||||||||
Density |
| ||||||||||||||||||||
Symmetry | Space group: 1 | ||||||||||||||||||||
Details | EMDB XML:
|
-Supplemental data
-Mask #1
File | emd_26461_msk_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: #1
File | emd_26461_half_map_1.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-Half map: #2
File | emd_26461_half_map_2.map | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Projections & Slices |
| ||||||||||||
Density Histograms |
-Sample components
-Entire : the rigor state wild type myosin-15-F-actin complex
Entire | Name: the rigor state wild type myosin-15-F-actin complex |
---|---|
Components |
|
-Supramolecule #1: the rigor state wild type myosin-15-F-actin complex
Supramolecule | Name: the rigor state wild type myosin-15-F-actin complex / type: complex / ID: 1 / Parent: 0 |
---|---|
Molecular weight | Theoretical: 5.4 kDa/nm |
-Supramolecule #2: Actin
Supramolecule | Name: Actin / type: complex / ID: 2 / Parent: 1 |
---|---|
Source (natural) | Organism: Gallus gallus (chicken) |
-Supramolecule #3: muscle regulatory light chain
Supramolecule | Name: muscle regulatory light chain / type: complex / ID: 3 / Parent: 1 |
---|---|
Source (natural) | Organism: Gallus gallus (chicken) |
-Supramolecule #4: Isoform 3 of Unconventional myosin-XV
Supramolecule | Name: Isoform 3 of Unconventional myosin-XV / type: complex / ID: 4 / Parent: 1 |
---|---|
Source (natural) | Organism: Mus musculus (house mouse) |
-Experimental details
-Structure determination
Method | cryo EM |
---|---|
Processing | single particle reconstruction |
Aggregation state | filament |
-Sample preparation
Buffer | pH: 7 Details: 10 mM imidazole pH 7.0, 50 mM KCl, 1 mM MgCl2, 1 mM EGTA, 0.5 mM DTT, 0.01% NaN3 |
---|---|
Grid | Model: C-flat-1.2/1.3 / Support film - Material: CARBON / Support film - topology: HOLEY |
Vitrification | Cryogen name: ETHANE / Chamber humidity: 100 % / Chamber temperature: 293 K / Instrument: LEICA EM GP |
Details | the ADP state wild type myosin-15 bound to F-actin |
-Electron microscopy
Microscope | FEI TITAN KRIOS |
---|---|
Image recording | Film or detector model: GATAN K2 SUMMIT (4k x 4k) / Detector mode: SUPER-RESOLUTION / Digitization - Frames/image: 1-40 / Number grids imaged: 1 / Average exposure time: 10.0 sec. / Average electron dose: 60.0 e/Å2 |
Electron beam | Acceleration voltage: 300 kV / Electron source: FIELD EMISSION GUN |
Electron optics | Illumination mode: FLOOD BEAM / Imaging mode: BRIGHT FIELD / Nominal defocus max: 3.5 µm / Nominal defocus min: 1.5 µm / Nominal magnification: 29000 |
Sample stage | Specimen holder model: FEI TITAN KRIOS AUTOGRID HOLDER / Cooling holder cryogen: NITROGEN |
Experimental equipment | Model: Titan Krios / Image courtesy: FEI Company |
+Image processing
-Atomic model buiding 1
Initial model | PDB ID: Chain - Source name: PDB / Chain - Initial model type: experimental model |
---|---|
Refinement | Space: REAL / Protocol: RIGID BODY FIT |